Xu Zhen, Yang Zujing, Bao Lisui, Lu Bei, Li Xiaoxu, Zhan Xin, Huang Xiaoting, Liu Yibing
Key Laboratory of Marine Genetics and Breeding (Ministry of Education), College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China.
Institute of Evolution & Marine Biodiversity, Ocean University of China, Qingdao 266003, China.
Antioxidants (Basel). 2024 Sep 4;13(9):1085. doi: 10.3390/antiox13091085.
Previous studies have shown that post-thaw sperm performance is affected by multiple stressors during cryopreservation, such as those induced by physical, chemical, mechanical and physiological changes. One of these is the balance disturbance between the antioxidant defense system and reactive oxygen species (ROS) production. This study investigated whether this disturbance could be alleviated by the addition of different antioxidants to cryoprotective solution [8% dimethyl sulfoxide (DMSO) in 1 µm filtered seawater] optimized for the sperm in dwarf surf clam , the model bivalve species used in many different types of studies. Results showed that the addition of 20 μM coenzyme Q10 (Q10) to 8% DMSO achieved a D-stage larval rate similar to that of the fresh control at a sperm-to-egg ratio at least 50% less than the 8% DMSO treatment alone. The addition of other antioxidants (glycine, melatonin and polyvinylpyrrolidone) did not have any positive effects. The improvement in post-thaw sperm quality by Q10 could be due to its ability to significantly decrease ROS production and lipid peroxidation and significantly increase the motility, plasma membrane integrity, mitochondrial membrane potential, acrosome integrity, DNA integrity and activities of catalase and glutatione. In this study, 37 fatty acids (FAs) were quantified in dwarf surf clam sperm, with 21 FAs being significantly impacted by the cryopreservation with 8% DMSO. Thirteen of these 21 FAs were changed due to the addition of 20 μM Q10 to 8% DMSO, with approximately half of them being improved significantly toward the levels of fresh control, while the remaining half extended further from the trends shown with 8% DMSO treatment. However, no significant difference was found in the percentage of each FA category sum and the ratio of unsaturated/saturated FAs between the two treated groups. In conclusion, the antioxidant Q10 has shown the potential to further improve the sperm cryopreservation technique in bivalves.
先前的研究表明，冷冻保存期间，解冻后精子的性能会受到多种应激源的影响，比如物理、化学、机械和生理变化所引发的应激源。其中之一是抗氧化防御系统与活性氧（ROS）生成之间的平衡紊乱。本研究调查了在为侏儒浪花蛤精子优化的冷冻保护溶液（1 µm过滤海水中的8%二甲亚砜（DMSO））中添加不同抗氧化剂，是否能够缓解这种紊乱，侏儒浪花蛤是许多不同类型研究中使用的双壳贝类模式物种。结果显示，在8% DMSO中添加20 μM辅酶Q10（Q10），在精子与卵子的比例至少比单独使用8% DMSO处理低50%的情况下，获得了与新鲜对照组相似的D期幼虫率。添加其他抗氧化剂（甘氨酸、褪黑素和聚乙烯吡咯烷酮）没有任何积极效果。Q10对解冻后精子质量的改善可能归因于其显著降低ROS生成和脂质过氧化的能力，以及显著提高活力、质膜完整性、线粒体膜电位、顶体完整性、DNA完整性以及过氧化氢酶和谷胱甘肽活性的能力。在本研究中，对侏儒浪花蛤精子中的37种脂肪酸（FAs）进行了定量，其中21种FAs受到8% DMSO冷冻保存的显著影响。在8% DMSO中添加20 μM Q10后，这21种FAs中有13种发生了变化，其中约一半显著改善至接近新鲜对照组的水平，而另一半则在8% DMSO处理所显示的趋势基础上进一步偏离。然而，两个处理组之间各FA类别总和的百分比以及不饱和/饱和FAs的比例没有显著差异。总之，抗氧化剂Q10已显示出进一步改进双壳贝类精子冷冻保存技术的潜力。
